EP0158057B1 - Salts of oxazaphosphorine derivatives - Google Patents

Description

beschrieben. In der vorstehenden Formel bedeuten :

• R_1' R₂ und R₃, die gleich oder verschieden voneinander sein können, Wasserstoff, Methyl, Ethyl, 2-Chlorethyl oder 2-Methansulfonyloxyethyl, wobei mindestens zwei dieser Reste 2-Chlorethyl und/oder 2-Methansulfonyloxyethyl sind,
• R₄ Wasserstoff oder Methyl,
• X ein gerad- oder verzweigtkettiges C_2-6-Alkylen, das eine Mercaptogruppe an dem 1-, 2-, 3-, 4- oder 5- ständigen Kohlenstoffatom der Alkylenkette haben kann, und
• Y⁸ das Wasserstoffkation, ein Alkali- oder Erdalkalikation, das Guanidinium-, Morpholinium- oder Cyclohexylammoniumkation oder das Kation, welches sich von einem Amin der Formel NR₅R₆R₇ ableitet, worin die Reste R₅ bis R₇ gleich oder verschieden sind und Wasserstoff, C₁-C₂-Alkylgruppen ⊕⊕ oder Oxyethylgruppen bedeuten, oder Y^⊕ ist das Ethylendiammoniumkation H₃N―CH₂CH₂―NH₃ oder das Piperazinoniumkation und z ist 1, wenn Y⁹ ein einbasisches Kation ist, oder z ist 2, wenn Y^⊕ ein zweibasisches Kation oder das Kation einer Verbindung mit zwei einbasischen Kationen ist.

In Belgian patent 892589 oxazaphosphorin-4-thio-alkanesulfonic acids and certain salts thereof are of the general formula

described. In the above formula:

• R _{1 '} R ₂ and R ₃ , which may be the same or different from one another, are hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl, at least two of these radicals being 2-chloroethyl and / or 2-methanesulfonyloxyethyl,
• R _{4 is} hydrogen or methyl,
• X is a straight or branched chain C _2-6 alkylene which can have a mercapto group on the 1-, 2-, 3-, 4- or 5-position carbon atom of the alkylene chain, and
• Y ^{8 is} the hydrogen cation, an alkali or alkaline earth metal cation, the guanidinium, morpholinium or cyclohexylammonium cation or the cation which is derived from an amine of the formula NR ₅ R ₆ R ₇ , in which the radicals R ₅ to R _{7 are} identical or different and hydrogen, C ₁ -C ₂ alkyl groups ⊕⊕ or oxyethyl groups, or Y ^⊕ is the ethylenediammonium cation H ₃ N ― CH ₂ CH ₂ ―NH ₃ or the piperazinonium cation and z is 1 if Y ^{9 is} a monobasic cation, or z is 2 if Y ^{⊕ is} a dibasic cation or the cation of a compound with two monobasic cations.

worin X Sauerstoff oder Schwefel ist, R₁, R₂ und R₃, die gleich oder verschieden voneinander sein können, Wasserstoff, Methyl, Ethyl, 2-Chlorethyl oder 2-Methansulfonyloxyethyl darstellen, die Reste R₄, die gleich oder verschieden voneinander sein können, Wasserstoff, Methyl oder Ethyl darstellen, R₅ Wasserstoff, C_1-4-Alkyl, Hydroxy-C_1-4-alkyl oder Phenyl ist, und Z unter anderem auch eine C₁-C₁₈-Alkylaminogruppe ist, die verschiedene Substituenten, darunter auch die Carboxygruppe enthalten kann und deren pharmazeutisch anwendbare Salze. Als Beispiel wird in dieser DE-OS eine Verbindung beschrieben worin Z die Gruppe ―NH―CH₂―CO₂H ist und das entsprechende Cyclohexylaminsalz.Furthermore, German Offenlegungsschrift 31 33 309 relates to 4-carbamoyloxy-oxazaphosphorin derivatives of the general formula

wherein X is oxygen or sulfur, R ₁ , R ₂ and R ₃ , which may be the same or different from each other, represent hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl, the radicals R ₄ which are the same or different from one another may represent hydrogen, methyl or ethyl, R _{5 is} hydrogen, C _1-4 alkyl, hydroxyC _1-4 alkyl or phenyl, and Z is, inter alia, also a C ₁ -C ₁₈ alkylamino group which has various substituents , including the carboxy group and their pharmaceutically acceptable salts. As an example, a compound is described in this DE-OS in which Z is the group ―NH ― CH ₂ ―CO ₂ H and the corresponding cyclohexylamine salt.

The compound of Belgian Patent 892589 and German Offenlegungsschrift 31 33309 have an anti-tumor effect and an immunosuppressive effect.

worin R₁, R₂ und R₃, die gleich oder verschieden voneinander sein können, Wasserstoff, Methyl, Ethyl, 2-Chlorethyl oder 2-Methansulfonyloxyethyl darstellen und dabei mindestens zwei dieser Reste 2-Chlorethyl und/oder 2-Methansulfonyloxyethyl sind und A die Gruppe -S-alk-S0₃H oder ―N(OH)―CONH―alk―CO₂H ist und alk einen, gegebenenfalls eine Mercaptogruppe enthaltenden, C₂-C₆-Alkylenrest darstellt, wobei alk auch -CH₂- sein kann, falls an der Gruppe alk die Carboxygruppe sitzt, mit Homocysteinthiolacton oder a-Amino-e-caprolactam oder einer basischen Verbindung der Formel

worin R₄ eine Hydroxygruppe, eine Aminogruppe oder eine C₁-C₆-Alkoxygruppe ist, R₅ Wasserstoff oder eine Difluormethylgruppe ist, R₆ Wasserstoff, einen Indolyl-(3)-methylrest, Imidazolyl-(4)-methylrest, eine C₁-C₁₀-Alkylgruppe oder eine C₁-C₁₀-Alkylgruppe, die durch eine Hydroxygruppe, eine C_l-C₆-Alkoxygruppe, eine Mercaptogruppe, eine C₁-C₆-Alkylthiogruppe, eine Phenylgruppe, eine Hydroxyphenylgruppe, eine Amino-C₁-C₆-alkylthiogruppe, eine Amino-C₁-C₆-alkyloxygruppe, eine Aminogruppe, eine Aminocarbonylgruppe, eine Ureidogruppe (H₂NCONH―), eine Guanidinogruppe oder eine C₁-C₆-Alkoxycarbonylgruppe substituiert ist, bedeutet oder worin R₆ zusammen mit dem StrukturteilThe invention relates to salts of oxazaphosphorin derivatives of the general formula

wherein R ₁ , R ₂ and R ₃ , which may be the same or different from each other, represent hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl and at least two of these radicals are 2-chloroethyl and / or 2-methanesulfonyloxyethyl and A the group -S-alk-S0 ₃ H or ―N (OH) ―CONH ― alk ― CO ₂ H and alk is a C ₂ -C ₆ alkylene radical which may contain a mercapto group, alk also -CH ₂ - if the carboxy group is located on the alk group, with homocysteine thiolactone or a-amino-e-caprolactam or a basic compound of the formula

wherein R _{4 is} a hydroxy group, an amino group or a C ₁ -C ₆ alkoxy group, R _{5 is} hydrogen or a difluoromethyl group, R _{6 is} hydrogen, an indolyl (3) methyl radical, imidazolyl (4) methyl radical, a C ₁ -C ₁₀ alkyl group or a C ₁ -C ₁₀ alkyl group through a hydroxy group, a C ₁ -C ₆ alkoxy group, a mercapto group, a C ₁ -C ₆ alkylthio group, a phenyl group, a hydroxyphenyl group, an amino -C ₁ -C ₆ alkylthio group, an amino-C ₁ -C ₆ alkyloxy group, an amino group, an aminocarbonyl group, an ureido group (H ₂ NCONH―), a guanidino group or a C ₁ -C ₆ alkoxycarbonyl group or wherein R ₆ together with the structural part

CR ₅ (NR ₇ R ₈ ) forms the proline residue, the 4-hydroxy-proline residue or the 2-oxo-3-amino-3-difluoromethyl-piperidine and the residues R ₇ and R _{8 are} hydrogen or C _i -C ₆ - Represent alkyl radicals.

• a) eine Verbindung der allgemeinen Formel
  
  worin X eine Hydroxygruppe oder eine C₁-C₄-Alkoxygruppe ist, mit dem Salz der Verbindung
  
  worin A die angegebenen Bedeutungen hat und die basische Salzkomponente Homocysteinthiolacton, α-Amino-ε-caprolactam oder die basische Verbindung der Formel II mit den angegebenen Bedeutungen für die Reste R₄ bis R₈ ist oder zuerst mit der Verbindung AH und anschließend mit den zuvor genannten basischen Verbindungen umsetzt oder eine Verbindung der allgemeinen Formel III, worin X eine gegebenenfalls duch eine Carboxygruppe, eine Hydroxygruppe oder C₁-C₄-Carbalkoxygruppe substituierte C₁-C₁₀-Alkylthiogruppe, eine Benzylthiogruppe oder eine C₁-C₆-Alkanoylthiogruppe ist, oder worin X die Gruppe ―N(OH)―CO―NHR darstellt und R Wasserstoff, eine C₁-C₆-Alkylgruppe, eine Benzylgruppe oder eine gegebenenfalls durch C₁-C₄-Alkylreste oder Halogen substituierte Phenylgruppe bedeutet, oder worin X die Gruppe A ist und der Rest X auch in der Salzform vorliegen kann, mit dem Überschuß einer Verbindung der Formel
  
  oder eines Salzes dieser Verbindung A'H oder mit Homocysteinthiolacton, α-Amino-ε-caprolactam oder der basischen Verbindung der Formel II mit den angegebenen Bedeutungen für die Reste R4 bis R₈ umsetzt, wobei A' von A verschieden ist und eine der für A angegebenen Bedeutungen hat oder
• b) eine Verbindung der Formel I oder das Salz einer Verbindung der allgemeinen Formel I mit Homocysteinthiolacton, a-Amino-e-caprolactam oder einer basischen Verbindung der Formel II mit den angegebenen Bedeutungen für R₄ bis R₈ beziehungsweise mit deren Salzen unter Bildung des entsprechenden Salzes umsetzt

und gegebenenfalls in den erhaltenen Verbindungen die basische Komponente oder den Säurewasserstoff der Gruppe A, falls diese nicht als Salz vorliegt, gegen eine andere basische Verbindung im Rahmen der hierfür gegebenen Definition austauscht.The. The invention further relates to a process for the preparation of salts of oxazaphosphorin derivatives of the formula I given above, which is characterized in that

• a) a compound of the general formula
  
  wherein X is a hydroxy group or a C ₁ -C ₄ alkoxy group with the salt of the compound
  
  wherein A has the meanings given and the basic salt component homocysteine thiolactone, α-amino-ε-caprolactam or the basic compound of formula II with the meanings given for the radicals R ₄ to R ₈ or first with the compound AH and then with the previously said basic compounds or a compound of the general formula III, wherein X is a C ₁ -C ₁₀ -alkylthio group optionally substituted by a carboxy group, a hydroxy group or a C ₁ -C ₄ -carbalkoxy group, a benzylthio group or a C ₁ -C ₆ -alkanoylthio group is or where X represents the group ―N (OH) ―CO ― NHR and R is hydrogen, a C ₁ -C ₆ alkyl group, a benzyl group or a phenyl group which is optionally substituted by C ₁ -C ₄ alkyl radicals or halogen, or wherein X is group A and the radical X can also be in the salt form with the excess of a compound of the formula
  
  or a salt of this compound A'H or with homocysteine thiolactone, α-amino-ε-caprolactam or the basic compound of the formula II is reacted with the meanings indicated for the radicals R 4 to R ₈ wherein A 'is different from A and one of the for A has the meanings given or
• b) a compound of formula I or the salt of a compound of general formula I with homocysteine thiolactone, a-amino-e-caprolactam or a basic compound of formula II with the meanings given for R ₄ to R ₈ or with their salts to form the corresponding salt

and, if appropriate, in the compounds obtained, the basic component or the Group A acid hydrogen, if this is not present as a salt, is replaced by another basic compound within the scope of the definition given for this.

The compounds according to the invention have a strong antitumor activity and can be used in particular to combat cancer. Compared to the previously known compounds of Belgian Patent 892 589 and German Offenlegungsschrift 31 33309, for example, they have reduced toxicity (for example reduced acute toxicity and leukotoxic activity) and therefore have an improved therapeutic index and better local and systemic and venous tolerance. Furthermore, the compounds according to the invention have a reduced immunosuppressive effect, a reduced local tissue irritation and often a lower hemolytic effect. Furthermore, the compounds according to the invention have no or only minor side effects on the circulatory system (for example sympathomimetic effects). Blood pressure is also less affected.

worin R₁, R₂, R₃ und A die hierfür angegebenen Bedeutungen haben. Gegenstand des Anspruchs 1 sind die Salze solcher Verbindungen der Formel I mit Homocysteinthiolacton oder a-Amino-e-caprolactam oder einer Verbindung der Formel II.The compounds of formula I have the following structure:

wherein R ₁ , R ₂ , R ₃ and A have the meanings given for this. The subject of claim 1 are the salts of such compounds of formula I with homocysteine thiolactone or a-amino-e-caprolactam or a compound of formula II.

insbesondere besteht die Kette alk aus 2 oder 3 C-Atomen, falls sie unverzweigt ist. Falls alk verzweigt ist, besteht der Teil, welcher an die Säuregruppe und an das Schwefel- beziehungsweise Stickstoffatom gebunden ist, insbesondere aus 2 oder 3 C-Atomen. Falls an der Gruppe alk die Carboxygruppe sitzt, ist alk vorzugsweise ―CH₂―. Falls der Rest alk eine Mercaptogruppe enthält (dies kann insbesondere der Fall sein, falls es sich um die Gruppe ―S―alk―SO₃H handelt), kann diese Mercaptogruppe an dem 1-, 2-, 3-, 4- oder 5-ständigen Kohlenstoffatom von alk sitzen. Die Zählung beginnt mit dem C-Atom an dem die saure Gruppe, zum Beispiel die -S0₃H-Gruppe sitzt. Insbesondere handelt es sich hierbei um den ―CH2―CH(SH)―CH₂-Rest.The alkylene radical (alk) of the formula I can be straight or branched. Examples of these are methylene, dimethylene, trimethylene, tetramethylene, pentamethylene or hexamethylene radical or, for example, the radicals or

in particular, the alk chain consists of 2 or 3 carbon atoms if it is unbranched. If alk is branched, the part which is bonded to the acid group and to the sulfur or nitrogen atom consists in particular of 2 or 3 carbon atoms. If the carboxy group is located on the alk group, alk is preferably ―CH ₂ -. If the rest alk contains a mercapto group (this can be the case in particular if it is the group ―S ― alk ― SO ₃ H), this mercapto group can be on the 1-, 2-, 3-, 4- or 5 - Sitting carbon atom from alk. The count begins with the C atom on which the acidic group, for example the -S0 ₃ H group, is located. In particular, this is the ―CH2 ― CH (SH) ―CH ₂ residue.

Such compounds are preferably suitable where R _{3 is} hydrogen, R ₁ and R _{2 are} each 2-chloroethyl and A is the group ―S ― CH ₂ ―CH ₂ ―SO ₃ H or ―N (OH) ―CO ―NH ― CH ₂ ―C0 ₂ H.

The alkyl groups, alkoxy groups, alkylthio groups occurring in formula II can be straight or branched. The C ₁ -C ₁₀ alkyl group preferably contains 1-6 carbon atoms. The alkoxy groups and alkylthio groups are preferably those with 1-4, in particular 1-2, carbon atoms; the same applies to the radicals R ₇ and R ₈ if these are alkyl radicals. If in formula II R _{6 is} an alkyl group which contains an amino-C ₁ -C ₆ -alkylthio- (C ₁ -C ₆ -alkoxy) group, then these are preferably the following groups: H ₂ N ― CH ₂ ―CH ₂ ―S ― CH ₂ - or H ₂ N ― CH ₂ ―CH ₂ ―O ― CH ₂ -

The compounds of the formula II are preferably those compounds where R _{4 is} a hydroxyl group and the radicals R ₅ , R ₇ and R _{8 are} hydrogen and R _{6 can} in particular take on the meanings given below. Preferred basic compounds of the formula II are, for example, those in which R _{4 is} a hydroxyl group, R _{5 is} hydrogen or difluoromethyl, R _{6 is} a C ₁ -C ₁₀ -alkyl group, in particular C ₁ -C ₆ -alkyl group, which (preferably in 2-, 3 -, 4-, 5- or 6-position; counting always begins at the point of connection of the alkyl radical with the Residual molecule) an amino group (in particular in the 3- or 4-position), an amino-C ₂ -C ₄ -alkylthio group, an amino-C ₂ -C ₄ -alkoxy group, a guanidino radical, an imidazolyl- (4) -methyl radical or one Contains indolyl- (3) -methyl and R ₇ and R _{B are} hydrogen or C ₁ -C ₄ -alkyl; or amino acid derivatives of the formula II, where R _{4 is} an amino group or a C ₁ -C ₄ alkoxy group, R _{5 is} hydrogen, R _{6 is} hydrogen, a phenylmethyl group, a 4-hydroxyphenylmethyl group or a C ₁ -C ₆ alkyl group, which (preferably in 2-, 3-, 4-, 5- or 6-position) contains a hydroxyl group, a mercapto group, a C ₁ -C ₄ alkylthio group, an aminocarbonyl group, a C ₁ -C ₄ alkoxycarbonyl group or ureido group and R ₇ and R _{8 is} hydrogen or C ₁ -C ₄ alkyl radicals.

In the case of the abovementioned amino acids or amino acid derivatives, the salts are each formed, for example, from one mole of oxazaphosphorin I and one mole of compound II.

Also suitable as basic compounds of the formula II are, for example, those where R _{4 is} an amino group or a C ₁ -C ₄ alkoxy group, R _{5 is} hydrogen or difluoromethyl, R _{6 is} a C ₁ -C 10 alkyl group, in particular C ₁ -C ₆ -alkyl group which (preferably in the 2-, 3-, 4- or w-position) an amino group, an amino-C ₂ -C ₄ -alkylthio group, an amino-C ₂ -C ₄ -alkoxy group, a guanidino radical, one Contains imidazolyl- (4) -methyl radical or an indolyl- (3) -methyl radical and R ₇ and R _{a are} hydrogen or C ₁ -C ₄ -alkyl radicals. In the last-mentioned amino acid derivatives, the salts are formed, for example, from 2 moles of oxazaphosphorine and 1 mole of the amino acid derivative of the formula II.

Individual examples of compounds of the formula II are: aspartic acid diamide (DL form), aspartic acid diethyl ester (L form), citrullinamide (H ₂ N ― CO ― NH- (CH ₂ ) ₃ ―CH (NH ₂ ) -CONH ₂ , L-form ), Ornithine ethyl ester (L-form), arginine, argininamide (L-form), 4-thialysine (H ₂ N ― CH ₂ ―CH ₂ ―S ― CH ₂ ―CH (NH ₂ ) ―COOH), 2,6- Diamino-enanthic acid (e-methyllysine), 4-oxalysine (H ₂ N ― CH ₂ CH ₂ ―O ― CH ₂ ―CH (NH ₂ ) ―COOH), glycinamide, N, N-dimethylglycinamide and the corresponding methyl or ethyl ester, prolinamide , Hydroxyprolinamide, phenylalaninamide, the methyl or ethyl ester of alanine or phenylalanine, homocysteine thiolactone (DL form), α-amino-ε-caprolactam (D (+) form)), lysine (especially L-lysine), difluoromethyl ornithine (DL or L form), valine methyl ester (L form), threonine ethyl ester, histidine methyl ester, histidinamide, alanine amide, ornithine.

In the salts according to the invention, in the event that R _{4 is} an amino group or an alkoxy group in the compound II and no other basic group is present or in the event that R _{4 is} the hydroxyl group and there is otherwise a basic group, Compound I (acid component) and compound II (basic component) practically in a ratio of 1: 1. (This also applies if the basic component is homocysteine thiolactone or α-amino-ε-caprolactam.) On the other hand, if the basic component II R _{4 is} an amino group or an alkoxy group and contains this in addition to the a-standing amino group, another basic group, then the ratio of compound I to compound II is generally 2: 1. The salts according to the invention are the neutral salts. In the case of sulfonic acid salts, for example, they have pH values between 3.5-6. If the salts are carboxylic acids, the pH values are, for example, between 6-9, in particular 6-8.

The procedure a)

The process is carried out in a solvent at temperatures between -60 ° C and + 90 ° C, preferably ―30 ° C to + 60 ° C, in particular -20 to + 30 ° C, that is, if necessary with cooling, at room temperature or below Heat. The reaction can be carried out in the presence of an acidic catalyst such as an inorganic or organic acid such as trichloroacetic acid. acid, p-toluenesulfonic acid, trifluoromethanesulfonic acid or a Lewis acid such as AlCl ₃ , ZnCl ₂ , TiCl ₄ , boron trifluoride etherate. For the reaction, for example, the pH is adjusted to between 1 and 8, preferably 2 and 6. This applies in particular when the starting components are used as salts, if appropriate also when the free acids are used and A contains in particular the carboxy group.

Examples of suitable solvents are: water, alcohols, in particular alkanols with 1-6 C atoms such as methanol, ethanol, propanol or isobutanol, alkyl ketones each with 1-4 C atoms such as in particular acetone, methyl ethyl ketone, aprotic solvents (for example Dimethyl sulfoxide, acetonitrile, N-methyl-pyrrolidone, dimethylformamide, hexamethylphosphoric acid triamide), halogenated hydrocarbons with 1-3 C atoms such as chloroform, ethylene dichloride, saturated cyclic ethers such as tetrahydrofuran, dioxane, saturated lower aliphatic ethers such as diethyl ether or similar solvents or mixtures of several such solvents.

If the symbol X in the starting compound III is the hydroxyl or alkoxy group, the reaction can also be carried out in two steps, for example by reacting the compound III first with the compound AH (without acidification) and then to the reaction mixture, if appropriate after concentration and addition of another of the solvents in question, the basic component II or homocysteine thiolactone or α-amino-ε-caprolactam is added in a solvent.

The use of a compound III, in which X is the hydroxyl group or an alkoxy group, is particularly suitable if the end products are obtained in crystalline form.

If X is not hydroxy or alkoxy, or if X is in the salt form, the compound becomes A'H or their salt is used in excess, for example 1.5-10 mol, preferably 2-5 mol, of the compound A'H or its salt per 1 mol of compound III. The pH of the reaction solution is adjusted, for example, to 5.5-9, preferably 6.5-8 using an alkali metal hydroxide solution or with an amine (expediently the amine which is already present as a basic component in the salt used); Under certain circumstances, a pH up to 12 can also be favorable. This applies in particular when the starting components are used as salts, if appropriate also when the free acids are used and A contains in particular the carboxy group. The reaction temperature is, for example, 10-90 ° C, preferably 25-60 ° C. The response time is, for example, a few seconds to several hours. The reaction solution is then, for example, quenched to below 10 ° C. and with a mineral acid (H ₂ SO ₄ , HCl, phosphoric acid), a sulfonic acid (for example mercapto-C ₁ -C ₆ alkanesulfonic acid) or an ion exchanger (H` form) brought to a pH between 4 to 5.5 or 7.

The products of the process can be isolated, for example: by crystallization or by a chromatographic process, in particular by preparative high-pressure liquid chromatography, if appropriate with subsequent conversion into the desired salt form on an appropriately loaded cation exchanger.

If X of formula III is group A and this is in the salt form, for example those salts are suitable which are described in German Offenlegungsschrift 31 33 309 or Belgian Patent 892589. For example, the ammonium salts, cyclohexylammonium salts or guanidinium salts are suitable. Of course, it is also possible to use other customary salts, for example optically active bases which are customary for resolution, which can be prepared analogously to the methods described therein.

The starting materials of the formula III are known, for example, from the following references or can be obtained analogously to the methods given therein: Belgian Patent 892 589, German Offenlegungsschrift 31 33 309, Tetrahedron Letters No. 10 (1979), pages 883-886, Cancer Treatment Reports, Volume 60, No. 4 (1976), pages 429-435. If X is an optionally substituted C ₁ -C ₁₀ -alkylthio group, compounds of the formula 111 are particularly suitable where X is a C ₁ -C ₆ -alkylthio group, the group ―S― (CH ₂ ) n ― CO ₂ H ( n = 1-6, especially 1-3), ―S― (CH ₂ ) _n ―OH (n = 2-6, especially 2-4) or ―S― (CH ₂ ) _n ―CO ― OC ₂ H ₅ (n = 1-6, especially 1-3). If X is a C ₁ -C ₆ -atkanoytthio group, it is in particular the acetylthio group. If X is the group ―N (OH) ―CO ― NHR and R is a C ₁ -C ₆ -alkyl group (straight or branched), this consists in particular of 1-4, preferably 1-2 C atoms.

The starting salts AH of formula IV or the starting salts A'H of formula V can be prepared, for example, by reacting a compound AH or A'H with a basic compound or homocysteine thiolactone or a-amino-e-caprolactam or a compound of formula II with or without solvents at temperatures between 0 and 40 ° C. Examples of suitable solvents are: water, C ₁ -C ₆ -alkanols (methanol, ethanol), lower aliphatic ketones (acetone), cyclic ethers (dioxane), chlorinated hydrocarbons (methylene chloride, ethylene dichloride, chloroform, carbon tetrachloride), saturated lower aliphatic ethers (diethyl ether), aprotic solvents (for example dimethylformamide, dimethyl sulfoxide, acetonitrile) or mixtures of these agents.

Such salts can also be prepared, for example, by dissolving an alkali salt (sodium salt) of the acid AH or A'H in water (for example 1 to 20% solution;% = percentages by weight), this solution through a column with a strongly acidic ion exchanger (H form, 3-fold excess) and the free acid in the eluate is neutralized with the basic component, concentrated in vacuo and, if necessary, the residue with a lower alcohol (methanol, ethanol), a lower ketone (acetone) or a Recrystallized ether (diethyl ether).

Procedure b)

This process is carried out at temperatures between 0-25 ° C, preferably 0-5 ° C. Examples of suitable solvents for this process are: water, lower aliphatic alcohols, lower aliphatic ketones or mixtures of these agents. 1 mole of component I is reacted with 1 mole of component II. It is advisable to work in a pH range between 3 and 8. When using compounds I, in which A is the S ― alk ― SO ₃ group (or a salt thereof), one preferably works at a pH between 3-6, preferably 3.8-5, in particular pH 4; when using compounds I in which A is the -N (OH) -CO-NH-alk-CO ₂ H group (or a salt thereof), the process is preferably carried out at pH values between 6-8, preferably pH 7 .

The addition of a buffer is often favorable. Examples of buffer systems with a pH range between 3.8 and 5.0 are: citric acid / sodium citrate; Acetic acid / sodium acetate; Phosphoric acid / sodium dihydrogen phosphate; Tartaric acid / sodium tartrate; Formic acid / sodium formate; Sodium hydrogen phosphate / citric acid; Succinic acid / amber sodium salt; Propionaries / sodium propionate; Aconitic acid / aconitic acid sodium salt; β, β-dimethylglutaric acid and its sodium salt; Maleic acid / sodium maleate; Compound II / citric acid. As a buffer for a pH range of 6 to 8 Examples include: Na citrate / NaOH, tris (hydroxymethyl) aminomethane maleate / NaOH, KH ₂ P0 ₄ / NaOH, KH ₂ ? 0 ₄ / Na ₂ HP0 ₄ .

Instead of the sodium hydroxide solution, homocysteine thiolactone or a-amino- _E -caprolactam or a basic compound of the formula II can be used to produce the buffers, so that the buffer already contains the basic component which is also present in the end product of the formula I.

The basic component of a salt of the compound I can be exchanged for a basic component according to the invention, for example on acidic ion exchangers which are loaded with homocysteine thiolactone, a-amino-e-caprolactam or the basic compound, II. In this case the basic compound II (which is now bound to the acid groups of the ion exchanger) is used in excess (for example 2 to 10 mol, preferably 5 mol of component II to 1 mol of component I). Examples of suitable acidic ion exchangers are those whose polymeric matrix contains sulfonic acid groups or carboxylic acid groups. The matrix of the ion exchanger can consist, for example, of a polystyrene resin, optionally with a content of 2 to 16, preferably 4 to 8, of divinylbenzene or a phenolic resin. The polystyrene ion exchanger is preferably in the form of a gel. The basic component can be charged to the ion exchanger in the following way, for example: 150 ml of ion exchange resin (1.2 meq / ml ^* ) in a column (diameter approx. 4 cm) with a cooling jacket are regenerated with hydrochloric acid, neutral with distilled water and free of chloride ions washed. The exchanger is then treated with a 10% aqueous solution of the basic compound (220 mmol) and washed neutral with distilled water. In addition, ion exchangers can be treated with a buffer (citric acid / citrate or acetic acid / acetate) of approximately pH 4 and then the buffer can be washed out again. Furthermore, the ion exchanger can also be loaded using neutral amino acid salts of the formula II or neutral salts of homocysteine thiolactone or a-amino-e-caprolactam (for example hydrochlorides or hydrobromides).

If ion exchangers are used, it is advantageous to add the buffer to the template for the eluate. In some cases, the elution of the salt together with the buffer and / or the salt of a mercapto-C ₂ -C ₆ -alkanesulfonic acid by the ion exchanger is also advantageous. It is also possible to elute the salt together with only the buffer. For example, the starting compound is dissolved, that is to say the compound I (A = ―S ― alk ― SO ₃ H) or a customary known salt of the compound I in a buffer at pH 3.8 to 5.0, preferably pH 4.1 and passes this solution over the ion exchange column and also collects the eluate in an appropriate buffer solution. The eluate or the lyophilisate produced therefrom then consists, for example, of the salt according to the invention and the buffer and / or the salt of mercaptoalkanesulfonic acid or of the salt according to the invention and only the buffer. The eluate is preferably immediately frozen or lyophilized, if appropriate after dilution with water. If the symbol A in the starting compound I is the group -N (OH) -CO-NH-alk-CO ₂ H, the procedure is expediently analogous, but now at pH values between 6-8 (for example pH 7).

However, one can also proceed in such a way that a customary salt of the compound I (for example alkali salt) in aqueous solution is passed through the acid form in the H form through an acidic ion exchanger as mentioned above and then the compound I in the eluate with the basic component of formula II or neutralized with homocysteine thiolactone or a-amino-e-caprolactam. This process option is particularly suitable if the end products are obtained in crystalline form.

Suitable starting salts of the compound of the formula I are, for example, those described in German Offenlegungsschrift 31 33 309 or Belgian Patent 892 589. For example, the ammonium salts, cyclohexylammonium salts or guanidinium salts are suitable. However, other customary salts (for example salts with optically active bases which are customary for the resolution of racemates), which can be prepared analogously to the methods described there, can also be used.

The oxazaphosphorine derivatives of the formula I according to the invention are understood to mean all possible stereoisomers and mixtures thereof. Specifically, these are, for example, cistrans isomers, that is to say the cis or trans position of group A to the oxo atom in the 2 position (phosphoryl oxygen) to the level of the oxazaphosphorine ring. For example, these are cis isomers and the trans isomers (in each case racemate and the corresponding enantiomers), the separate cis isomers and the separate trans isomers. Diastereomeric salts (for example if a chiral amine is used for salt formation) can preferably be separated in a known manner by fractional crystallization. The pure enantiomers can be obtained by conventional methods of resolving racemates, for example by fractional crystallization of the diasteromeric salts from racemic acids of the formula I and optically active bases or, if appropriate, by using optically active starting materials of the formula III in the synthesis.

In general, cis / trans mixtures can arise in the synthesis. In general, mixtures are formed which consist predominantly of the cis isomer and approximately 5-10% of the trans isomer.

^* The manufacturers of the ion exchangers state the capacity of the ion exchangers (ie the amount of functional groups such as -S0 ₃ H, ―CO ₂ H) in meq / ml or meq / g of the ion exchange resin.

The compounds according to Examples 1-5 consist, for example, of the cis isomer with less than 5-10% of the trans form.

In the case of compounds which crystallize well, the cis or trans form, in particular the cis form, is crystallized from such mixtures. However, if the reaction is carried out in anhydrous solvents or in solvents with a low water content, only one form, or in particular the cis form, is obtained. For example, the pure cis form of a compound of formula I which does not crystallize or has poor crystallization can be prepared by adding an acetone solution of the compound of formula 111 to an aqueous solution of the compound of formula IV or its salt at temperatures between -30 and + 20 ° C is given and is reprecipitated several times after the reaction has ended.

The starting compounds according to formula 111 can be used as racemic cis and trans isomers (see illustration on the previous page), as optically active cis and trans form and as mixtures thereof (see Belgian patent 892 589 and German Offenlegungsschrift 31 33 309, Page 12).

For example, 1-phenylethylamine, brucine, quinidine, strychnine and cinchonine as well as further bases and methods come into question for the resolution of racemates as well as further bases and methods, which are described in “Optical Resolution Procedures for Chemicals Compounds •, Vol. 2, Paul Newman, 1981, Verlag Optical Resolution Information Center in Riverdale, USA. For this purpose, for example, a racemic salt according to the invention is converted in the manner already specified into a salt with one of the optically active bases mentioned above, the enantiomers are separated in a known manner and the optically active base of the enantiomers thus obtained is then replaced by a basic compound according to Invention. However, the above-mentioned optically active bases can also be used as a basic salt component in the synthesis in process a) in the reaction of a compound of formula III with a compound of formula IV or V or in process b). In this case, this optically active base must then be exchanged in the usual way for the basic salt component according to the invention in accordance with the definition already given.

The basic salt components homocysteine thiolactone, α-amino-ε-caprolactam according to the invention and the compounds of the formula 11 are suitable as racemates or in the form of the pure enantiomers.

In general, the L-forms are preferred.

The salts according to the invention are understood to mean all forms which result from the various asymmetric carbon atoms present, for example racemates, optically active forms, diastereomeric forms.

During production, it is advisable to keep the salts according to the invention as short as possible in solution in order to hydrolysis to 4-hydroxyoxazaphosphorines and / or epimerization on the C-4 atom of the oxazaphosphorine ring (conversion of the cis form into the trans form) to prevent or keep it as low as possible. If the salts according to the invention are more contaminated (for example contain larger amounts of starting compounds), they can be obtained in pure form by customary chromatographic methods (in particular by preparative high-pressure liquid chromatography).

The salts according to the invention are stable, have a shelf life (in particular at 4 ° C.) and can be processed galenically well. For galenical preparations (for example hydrolysis-stable injection solutions, lyophilisates), in particular also with regard to storage stability, it is advisable to set a pH range of approx. 3.5-7 for the sulfonic acid derivatives using a conventional buffer (e.g. citrate buffer). The pH optimum is pH 4.0-4.3. In the event that the radical A is derived from the group -N (OH) -CO-NH-alk-CO2H, a pH of 6.5-7.5 is expediently set. These pH value adjustments can be made for solutions and suspensions as well as for solid pharmaceutical preparations.

In addition and regardless of the addition of a buffer, the addition of 0.5 to 5 equivalents of a salt (for example an alkali salt, in particular sodium salt) of a mercapto-C ₂ -C ₆ -alkylsulfonic acid (for example a salt of 2-mercapto-ethanesulfonic acid) ) and their disulfides and other thiols (for example cysteine) advantageous. The type of thiols and disulfides and the mode of their use are described in European patent application 83439. Examples of such salts are the alkali metal salts (Na, K) or the salts with a basic component according to the invention (compound of the formula 11, homocysteine thiolactone, a-amino- _E- caprolactam).

The addition of the salt of a mercapto-C ₂ -C _a -alkanesulfonic acid can, for example, by adding an aqueous solution of the sulfonic acid salt (alkali salt, for example 20 percent by weight) to an aqueous solution of the salt according to the invention (preferably buffered to a pH between 4 and 4.3 ) respectively. The mixture thus obtained is then lyophilized, for example.

The advantages of adding a mercaptoalkanesulfonic acid salt in this way are as follows: improvement of the storage stability and the stability in aqueous solution in the case of the salts according to the invention. (This is important, for example, in the preparation of the salts but also in the dissolution of, for example, lyophilizates before use); Improvement in chemotherapy of cancer by means of the salts according to the invention, in particular with regard to toxic side effects (see European patent application 83 439, European patent 2495, German patent 28 06 866).

The salts according to the invention are suitable for the production of pharmaceutical compositions or preparations. The pharmaceutical compositions or pharmaceuticals contain as active ingredient one or more of the salts according to the invention, optionally in a mixture with other pharmacologically or pharmaceutically active substances. The pharmaceuticals can be produced using the known and customary pharmaceutical carriers and auxiliaries.

The pharmaceutical compositions may, for example, enteral, parenteral, oral, perlingually or applied in the form of _sprays'. The administration can take place, for example, in the form of tablets, capsules, pills, dragées, suppositories, liquids or aerosols. Examples of possible liquids are: oily or aqueous solutions or suspensions, emulsions, injectable aqueous or oily solutions or suspensions.

When administered intravenously, intraperitoneally or orally, the compounds according to the invention have good cytostatic and curative activity in various experimental tumors of rats and mice. For example, the compounds according to the invention are administered to the rat 5 days after intraperitoneal implantation of 10 ⁵ cells of leukemia L5222 with different doses intravenously, intraperitoneally or orally and, depending on the dose, a curative effect is achieved. The recurrence and metastasis-free survival of the tumor-bearing animals after 90 days is defined as the cure. The dose with which 50% of the tumor-bearing animals can be cured is calculated from the frequency of healings obtained with the different doses by means of probit analysis according to R. Fisher as the mean curative dose (DC50).

For example, the compounds according to the invention are also administered one day after intraperitoneal implantation of 10 ⁶ cells of Yoshida ascites sarcoma AH13 with different doses intravenously, intraperitoneally or orally and a curative effect is achieved depending on the dose.

Here too, the curative effect is defined as relapse and metastasis-free survival of the tumor-bearing animals over 90 days.

Correspondingly, Probit analysis according to R. Fisher calculates the mean curative dose (DC50) at which 50% of the tumor-bearing animals can be cured.

Furthermore, for example, the compounds according to the invention are administered once or several times (4 ×) on successive days after intraperitoneal implantation of 10 ^a cells of mouse leukemia L1210 with different doses intravenously, intraperitoneally or orally and a cytostatic effect is achieved.

The cytostatic effectiveness is to be recorded as an extension of the median survival time of the tumor animals and is expressed as a dose-dependent percentage increase in survival time compared to an untreated control group.

The mean curative dose for rat tumors is in the range of 0.1-10 mg / kg regardless of the dosage form. With the same doses, a median survival extension of 100% for L1210 mouse leukemia can be achieved.

In addition, the compounds according to the invention stimulate antibody production in a certain low dose range. For the compound according to Example 5, this dose range is, for example, between 20-50 mg / kg rat (intravenous, intraperitoneal). In contrast, antibody production in the same dose range is already suppressed by the known anti-tumor agent cyclophosphamide.

• N. Brock : Pharmakologische Grundlagen der Krebs-Chemotherapie.

Literature:

• N. Brock: Pharmacological foundations of cancer chemotherapy.

In: A. Georgii (ed.), Negotiations of the German Cancer Society Volume 1, pp. 15-42, Gustav Fischer Verlag, Stuttgart (1978).

This curative and cytostatic effect is comparable to the effects of the well-known drugs cyclophosphamide and ifosfamide.

• 0,01 mg/kg oral
• 0,01 mg/kg intraperitoneal
• 0,01 mg/kg intravenös.

The lowest, already curative or cytostatic effective dose in the animal experiments given is, for example

• 0.01 mg / kg orally
• 0.01 mg / kg intraperitoneally
• 0.01 mg / kg intravenously.

• 0,01-100 mg/kg oral, insbesondere 0,1-10,0 mg/kg
• 0,01-100 mg/kg intraperitoneal, insbesondere 0,1-10,0 mg/kg
• 0,01-100 mg/kg intravenös, insbesondere 0,1-10,0 mg/kg.

As a general dose range for the curative and cytostatic effect (animal experiment as above) there are for example:

• 0.01-100 mg / kg orally, in particular 0.1-10.0 mg / kg
• 0.01-100 mg / kg intraperitoneally, in particular 0.1-10.0 mg / kg
• 0.01-100 mg / kg intravenously, especially 0.1-10.0 mg / kg.

Indications for which the compounds according to the invention can be considered: malignant diseases in humans and animals.

The pharmaceutical preparations generally contain between 1 mg to 1 g, preferably 100 to 1,000 mg, of the active component (s) according to the invention.

The administration can take place, for example, in the form of tablets, capsules, pills, dragées, suppositories, ointments, jellies, creams, powders, dusting powder, aerosols or in liquid form. Possible liquid application forms are, for example: oily or alcoholic or aqueous solutions as well as suspensions and emulsions. Preferred forms of use are tablets which contain between 10 and 200 mg or solutions which contain between 0.1 and 5% of active substance.

• a) bei oralen Arzneiformen zwischen 1-100 mg/kg, vorzugsweise 10-60 mg/kg,
• b) bei parenteralen Arzneiformen (zum Beispiel intravenös, intramuskulär) zwischen 1-100 mg/kg, vorzugsweise 10-60 mg/kg,
• c) bei Arzneiformen zur rektalen oder vaginalen Applikation zwischen 1-100 mg/kg, vorzugsweise 10-60 mg/kg,
• d) bei Arzneiformen zur lokalen Applikation auf die Haut und Schleimhäute (zum Beispiel in Form von Lösungen, Lotionen, Emulsionen, Salben und so weiter) zwischen 1-100 mg/kg, vorzugsweise 10-60 mg/kg.

The single dose of the active components according to the invention can be, for example

• a) in the case of oral dosage forms between 1-100 mg / kg, preferably 10-60 mg / kg,
• b) in the case of parenteral pharmaceutical forms (for example intravenous, intramuscular) between 1-100 mg / kg, preferably 10-60 mg / kg,
• c) in the case of pharmaceutical forms for rectal or vaginal application between 1-100 mg / kg, preferably 10-60 mg / kg,
• d) in the case of pharmaceutical forms for local application to the skin and mucous membranes (for example in the form of solutions, lotions, emulsions, ointments and so on) between 1-100 mg / kg, preferably 10-60 mg / kg.

The doses are based on the free base.

For example, 1 to 10 tablets containing 10 to 300 mg of active substance 1-3 times a day or, for example, in the case of intravenous injection 1 or 2 times a day, one or more ampoules of 1 to 10 ml content containing 10 to 250 mg of substance are recommended . For oral administration, the minimum daily dose is, for example, 200 mg; the maximum daily dose for oral administration should not exceed 5,000 mg.

A continuous infusion of appropriate doses over 12 and more hours can also be recommended in individual cases.

For the treatment of dogs and cats, the single oral dose is generally between about 10 and 60 mg / kg body weight; the parenteral dose is approximately between 10 and 60 mg / kg body weight.

For the treatment of horses and cattle, the single oral dose is generally between about 10 and 60 mg / kg body weight; the parenteral single dose approximately between 10 and 60 mg / kg body weight.

The acute toxicity of the compounds according to the invention on the mouse (expressed by the LD 50 mg / kg; method according to Miller and Tainter: Proc. Soc. Exper. Biol. A. Med. 57 (1944) 261) is, for example, between 100 when administered orally and 1,000 mg / kg (or above 1,000 mg / kg).

The medicinal products can be used in human medicine, veterinary medicine and agriculture alone or in a mixture with other pharmacologically active substances.

example 1 4- (2-sulfo-ethylthio) cyclophosphamide ^* glycine amide salt

4.0 g (18 mmol) of 4-hydroxycyclophosphamide in 10 ml of distilled water are mixed with 5 g (18 mmol) of 2-mercaptoethanesulfonic acid glycine amide salt in 40 ml of acetone. The reaction solution is acidified with trichloroacetic acid to a pH of 4, kept at 5 ° C. for 2 hours and at -25 ° C. for 20 hours. The salt separates out in crystalline form and is filtered off, washed, dried and recrystallized from water / acetone.

F. 90-98 ° C; Yield: 7.2 g (76% of theory). The salt contains about 1 equivalent of acetone.

Example 2 4- [1-hydroxy-3-carboxymethyl-ureido (1)] cyclophosphamide lysine salt (L-lysine)

7.5 g (24.6 mmol) of 4-ethoxycyclophosphamide and 3 g (22.4 mmol) of 1-hydroxy-3-carboxymethyl urea are stored in 50 ml of dry alcohol for 20 hours at 0 ° C. The reaction solution is then concentrated in vacuo at 20 ° C., the residue is taken up in 200 ml of acetone, 3.3 g (22.6 mmol) of L-lysine in 25 ml of methanol are added and the gelatinous precipitate is centrifuged after standing for a short time. The residue is dissolved in water, filtered, precipitated with acetone, suction filtered and washed with acetone / ether.

Mp 125-128 ° C; Yield: 6.5 g (54% of theory).

^* Cyclophosphamide = 2- [2- (bis (2-chloroethyl) amino) -2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorine.

Example 3 4- (2-sulfo-ethylthio) cyclophosphamide glycinamide salt.

A solution of 3.1 g (6.5 mmol) of 4- (3-sulfopropylthio) cyclophosphamide guanidine salt and 4.2 g (19.6 mmol) of 2-mercaptoethanesulfonic acid glycinamide salt in 15 ml of water is adjusted to pH 7 with sodium hydroxide solution , 5 and heated to about 40 ° C for 5 minutes. The reaction solution is then quenched to 0 ° C., adjusted to pH 4.5 with sulfuric acid, mixed with 70 ml of acetone and stored at 4 ° C. for 3 days. The precipitate is filtered off and recrystallized from water / acetone.

F. from 85 ° C (decomposition); Yield: 430 mg (14% of theory).

Example 4 4- (2-sulfo-ethylthio) cyclophosphamide arginine salt (L-arginine).

37.2 g (74.5 mmol) of 4- (2-sulfo-ethylthio) -cyclophosphamide-cyclohexylamine salt are dissolved in 320 ml of distilled water at 5 ° C. (pH 4.3) and passed through a column cooled to 4 ° C. Given 150 ml of cation exchanger. It is a gel-like polystyrene resin with a content of 8% divinylbenzene, which contains sulfonic acid groups that are loaded with arginine.

Flow rate: 6 ml / minute. Rinse with 250 ml of water. The eluate cooled to 4 ° C is diluted with cold water to a 5% solution and then lyophilized.

F. 85-90 ° C (decomposed); Yield: 42.9 g (100% of theory).

A buffer substance and / or a mercaptoalkylsulfonic acid can be added as follows:

a) Addition of sodium citrate buffer

The procedure is as described above, but the eluate is collected in sodium citrate buffer (pH 4.1) and subsequently diluted, for example, to a solution which is 0.5 molar in sodium citrate and 5% in active ingredient. Finally, it is lyophilized.

Yield: 42.9 g (100% of theory), compound according to Example 4 with sodium citrate buffer.

b) Add sodium citrate buffer and sodium 2-mercapto-ethanesulfate _Q nat (Mesna).

The procedure is as described above, but the eluate is collected in a cooled sodium citrate buffer solution (pH 4.1), which contains 8.2 g (50 mmol) of mesna, and then lyophilized.

Yield: 42.9 g (100% of theory) of compound according to Example 4 with mesna and sodium citrate buffer.

Example 5 4- (2-sulfo-ethylthio) cyclophosphamide lysine salt (L-lysine).

37.3 g (74.5 mmol) of 4- (2-sulfo-ethylthio) -cyclophosphamide-cyclohexylamine salt are eluted analogously to Example 4 on an ion exchanger (protonated lysine form) and then lyophilized.

F. from 85 ° C (decomposition); Yield: 40.8 g (100% of theory).

A changed way of working is for example the following:

82.4 g (16.5 mmol) of 4- (2-sulfo-ethylthio) cyclophosphamide cyclohexylamine salt are dissolved in 850 ml of distilled water at 5 ° C. The solution is adjusted to pH 4.1 with a few grains of strongly acidic ion exchanger (with sulfonic acid groups) and at a dropping rate of 50 ml / minute over a column cooled to 4 ° C. with 800 ml ion exchanger ^* with 820 mmol sulfonic acid L-lysine groups given. The first 180 ml of the eluate are discarded and the subsequent eluate is continuously adjusted to pH 4.1 with a total of about 1.5 ml of strongly acidic ion exchanger (with sulfonic acid groups) while stirring and cooling with ice water. The ion exchanger is rinsed with 900 ml of distilled water cooled to 0 ° C. The eluate is then diluted with cold water to a 5% solution and immediately lyophilized.

F. from 85 ° C, yield: 90 g (100% of theory).

Compound according to Example 5 with lysine citrate buffer (lysine + citric acid).

7.2 g (14.4 mmol) of 4- (2-sulfo-ethylthio) cyclophosphamide cyclohexylamine salt are dissolved in 80 ml of 0.05 M sodium citrate buffer (pH 4.1) at 4 ° C. and over a temperature of 4 ° C. cooled column with 40 ml of cation exchange resin (protonated lysine form). It is rinsed with 80 ml of buffer solution, the eluate is made up to 150 g of solution and lyophilized.

Yield: 7.9 g (100% of theory) of compound according to Example 5 with lysine citrate buffer.

^* It is the same ion exchanger as in example 4.

Compound according to Example 5 with 2-mercaptoethanesulfonic acid lysine salt and lysine citrate buffer.

7.2 g (14.4 mmol) of 4- (2-sulfo-ethylthio) cyclophosphamide cyclohexylamine salt and 1.5 g (9.2 mmol) of mesna are dissolved in 80 ml of 0.05 M sodium citrate buffer pH 4.1 at 4 ° C and passed through a column cooled to 4 ° C with 40 ml of cation exchange resin (protonated lysine form). It is rinsed with 50 and 30 ml of buffer solution, the cooled eluate is made up to 150 g of solution and lyophilized.

Yield: 7.9 g (100% of theory) of the compound according to Example 5 with 2.6 g (100% of theory) of 2-mercaptoethanesulfonic acid lysine salt and lysine citrate buffer (pH 4.1).

Example 6 4- (2-sulfoethylthio) cyclophosphamide glycinamide salt.

2.9 g (8.6 mmol) of 4- (2-hydroxyethylthio) cyclophosphamide and 5.6 g (26 mmol) of 2-mercaptoethanesulfonic acid glycinamide salt are dissolved in 35 ml of distilled water. The pH is adjusted to about 8 with glycinamide. The reaction solution is heated to about 40 ° C. for about 4 minutes, then quenched to 0 ° C. and, after the pH has been lowered to about 4.5 by means of 2-mercapto-ethanesulfonic acid, 500 ml of acetone are added. After a few days at 4 ° C., the product is filtered off with suction and recrystallized from water / acetone.

F. from 85 ° C.

Yield: 410 mg (9% of theory).

The salt contains about 1 equivalent of acetone.

Examples of pharmaceutical preparations a) Example of the preparation of a lyophilizate of 4-sulfo-ethylthio-cyclophosphamide lysine salt.

90 g of 4-sulfo-ethylthio-cyclophosphamide L-lysine salt are dissolved in 1,500 ml of water cooled to 4 ° C. for injection purposes with stirring. The mixture is then made up to 1800 ml with water from 4 ° C. With a few grains of regenerated cation exchanger (H form) the pH is adjusted to about 4.2. To prepare the lyophilisates, the above solution is subjected to sterile filtration in a known manner. The collecting vessel is cooled. All operations following sterile filtration are carried out under aseptic conditions.

The sterile solution is filled to a volume of 2 ml in a 10 ml injection bottle. The active substance content is 100 mg.

The bottles are provided with sterile freeze-drying stoppers and lyophilized in a freeze-drying system. The system is then aerated with sterile, dried nitrogen and the ampoule bottles in the system are closed. To prepare an applicable injection solution, the contents of the bottles are dissolved in 5 ml of water for injections.

The lyophilisate should be stored at 0-6 ° C (refrigerator).

b) Example of the preparation of a lyophilizate of 4- (2-sulfo-ethylthio) cyclophosphamide lysine salt (L-lysine) with citric acid-lysine buffer.

• 1. An ion exchange column with a cooling jacket is loaded with 1 300 ml of an acidic ion exchanger, regenerated with 2 liters of 10% hydrochloric acid and washed neutral and free of chloride ions with water for injection purposes.
• 2. The column is then loaded with 3 liters of a 10% lysine solution, freed from excess lysine by washing with water for injection purposes and washed until neutral.
• 3. 1.4 liters of a solution of the following composition are passed over the column:
  

Active ingredients and auxiliary substances are dissolved in water at approx. 4 ° C. The pH of the solution is 4.1. The cation exchange column is now also cooled to approx. 4 ° C. The above solution is given over the column. The flow rate is 10 ml / minute.

The eluate is collected in a cooled template, the first 300 ml being discarded as a preliminary. The column is then washed with water cooled to 4 ° C. for injection purposes and the total volume of the eluate is made up to 3 liters.

The eluate to be processed into lyophilisates has the following composition:

4. To prepare the lyophilisates, the above solution is subjected to sterile filtration in a known manner. The collecting vessel is cooled. All operations following sterile filtration are carried out under aseptic conditions.

• 2 ml Lösung in eine 10 ml Injektionsflasche. Der Wirkstoffgehalt beträgt 100 mg.
• 10 ml Lösung in eine 30 ml Injektionsflasche. Der Wirkstoffgehalt beträgt 500 mg.

The sterile solution is filled into injection bottles as follows:

• 2 ml solution in a 10 ml injection bottle. The active substance content is 100 mg.
• 10 ml solution in a 30 ml injection bottle. The active substance content is 500 mg.

The bottles are provided with sterile freeze-drying stoppers and lyophilized in a freeze-drying system. The system is then aerated with sterile, dried nitrogen and the ampoule bottles in the system are closed.

To prepare an applicable injection solution, the contents of the bottles with 100 mg of active ingredient in 5 ml and with 500 mg of active ingredient in 25 ml of water are dissolved for injection purposes.

c) Example of the preparation of a lyophilizate of 4-sulfo-ethylthio-cyclophosphamide arginine salt.

90 g of 4-sulfo-ethylthio-cyclophosphamide arginine salt and 135 g of sodium 2-mercapto-ethanesulfonate are dissolved in 1,500 ml of water cooled to 4 ° C. for injection purposes with stirring. After complete dissolution, make up to 1800 ml with water from 4 ° C. With a few grains of regenerated cation exchanger (H ⁺ form) the pH is adjusted to about 4.2. To prepare the lyophilisates, the above solution is subjected to sterile filtration in a known manner. The collecting vessel is cooled. All operations following sterile filtration are carried out under aseptic conditions. The sterile solution is filled to a volume of 2 ml in a 10 ml injection bottle. The active substance content is 100 mg. The bottles are provided with sterile freeze-drying stoppers and lyophilized in a freeze-drying facility. The system is then aerated with sterile, dried nitrogen and the ampoule bottles in the system are closed. To prepare an applicable solution for injection, the contents of the bottles are dissolved in 5 ml of water for injections.

The lyophilisate should be stored at 0-6 ° C (refrigerator).

Claims (12)

1. Salts of oxazaphosphorine derivatives of the general formula

wherein R₁, R₂ and R₃, which can be identical or different from one another, represent hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulphonyloxyethyl and in this case at least two of these radicals are 2-chloroethyl and/or 2-methanesulphonyloxyethyl and A is the group ―S―alk―SO₃H or ―N(OH)―CONH―alk―CO₂H and alk represents a C₂-C₆-alkylene radical optionally containing a mercapto group, where alk can also be -CH₂- if the carboxyl group is on the group alk, with homocysteine thiolactone or α-amino-ε-caprolactam or a basic compound of the formula

wherein R₄ is a hydroxyl group, an amino group or a C_l-C₆-alkoxy group, R₅ is hydrogen or a difluoromethyl group, R₆ denotes hydrogen, a 3-indolylmethyl radical, a 4-imidazolylmethyl radical, a C₁-C₁₀-alkyl group or a C₁-C₁₀-alkyl group which is substituted by a hydroxyl group, a C₁-C₆-alkoxy group, a mercapto group, a C_l-C₆-alkylthio group, a phenyl group, a hydroxyphenyl group, an amino-C_l-C₆- alkylthio group, an amino-C₁-C₆-alkyloxy group, an amino group, an aminocarbonyl group, a ureido group (H₂NCONH―), a guanidino group or a C_l-C₆-alkoxy-carbonyl group, or wherein R₆ together with the structural moiety CR₅(NR₇R₈) forms the proline radical, the 4-hydroxy-proline radical or the 2-oxo-3-amino-3-difluoromethylpiperidine and the radicals R₇ and R_a represent hydrogen or C₁-C₆-alkyl radicals.

2. Salts according to Claim 1, characterized in that the basic compound is a compound of the formula II, wherein R₄ is a hydroxyl group or the amino group, the radicals R₅, R₇ and R₈ are hydrogen and R₆ denotes hydrogen or a C₁-C₄-alkyl group which, in particular in the w-position, can be substituted by an amino group or a guanidino group.

3. Process for the preparation of salts of oxazaphosphorine derivatives of the general formula I according to Claim 1, characterized in that a compound of the general formula

wherein X is a hydroxyl group or a C₁-C₄-alkoxy group, is reacted with the salt of the compound

wherein A has the meanings indicated and the basic salt component is homocysteine thiolactone, a-amino-ε-caprolactam or the basic compound of the formula II having the meanings indicated for the radicals R₄ to R_B.

4. Process for the preparation of salts of oxazaphosphorine derivatives of the general formula I according to Claim 1, characterized in that a compound of the general formula

wherein X is a hydroxyl group or a C₁-C₄-alkoxy group, is first reacted with the compound

and subsequently with homocysteine thiolactone, a-amino-e-caprolactam or the basic compound of the formula II.

5. Process for the preparation of salts of oxazaphosphorine derivatives of the general formula I according to Claim 1, characterized in that a compound of the general formula III, wherein X is a C₁-C₁₀- alkyl-thio group which is optionally substituted by a carboxyl group, a hydroxyl group or a C,-C₄- carbalkoxy group, a benzylthio group or a C₁-C₆-alkanoylthio group, or wherein X represents the group ―N(OH)―CO―NHR and R denotes hydrogen, a C₁-C₆-alkyl group, a benzyl group or a phenyl group which is optionally substituted by C,-C₄-alkyl radicals or halogen, or wherein X is the group A and the radical X can also be present in the salt form, is reacted with excess of a compound of the formula

or a salt of this compound A'H or with homocysteine thiolactone, a-amino-_B-caprolactam or the basic compound of the formula II having the meanings indicated for the radicals R₄ to R₈, where A' is different from A and has one of the meanings indicated for A.

6. Process for the preparation of salts of oxazaphosphorine derivatives of the general formula I according to Claim 1, characterized in that a compound of the formula I or the salt of a compound of the general formula I is reacted with homocysteine thiolactone, a-amino-e-caprolactam or a basic compound of the formula II having the meanings indicated for R₄ to R₈ or with their salts with the formation of the corresponding salts.

7. Process for the preparation of salts of oxazaphosphorine derivatives of the general formula I according to Claim 1, characterized in that the basic components or the acidic hydrogen of the group A, if this is not present as a salt, is exchanged for another basic compound in the context of the definition given therefor in compounds of the formula I.

8. Medicaments, containing a salt according to one or more of the foregoing claims in addition to customary excipients and/or diluents or auxiliaries.

9. Medicaments according to Claim 8, characterized in that it additionally contains a buffer and/or the alkali metal salt of a mercapto-C₂-C₆-alkanesulphonic acid.

10. Process for the preparation of a medicament, characterized in that a compound according to Claim 1 or 2 is processed to give pharmaceutical preparations using customary pharmaceutical excipients or diluents or other auxiliaries, or is brought into a therapeutically utilizable form.

11. Process according to Claim 10, characterized in that buffer and/or the alkali metal salt of a mercapto-C₂-C_a-alkanesulphonic acid are additionally used.

12. Use of compounds according to Claim 1 or 2 for the preparation of medicaments.